Genetic safety and robustness of base editing and simultaneous hiPSC generation
(A) PCR to detect episomal vector retention into four hiPSC lines reprogrammed in absence of the ABEmax construct (−ABE) and eight independently base-edited hiPSC lines (+ABE). Two different plasmid regions are targeted in these PCRs: EBNA-1 and OriP. Water control (−), positive control (+), NOTCH3: patient 1 (Pt 1) and patient 2 (Pt 2). LDLR: patient 3 (Pt 3) and patient 4 (Pt 4).
(B) Karyotyping of a representative base-edited hiPSC lines and its non-ABE control.
(C) Bar graphs depicting the A-to-G editing frequency on the targeted loci: gRNA2 (chr11:-5254881) and Site 16 (chr1:-179826685). Each point represents an independent event of simultaneous reprogramming and base editing on primary fibroblasts derived from five different donors (n = 5). Data are represented as mean + standard deviation.